BACKGROUND Quinine oxidoreductase 1(NQO1)plays a vital role in protecting normal cells against oxidative damage and electrophilic attack.It is highly expressed in many solid tumors,suggesting a role in cancer developm...BACKGROUND Quinine oxidoreductase 1(NQO1)plays a vital role in protecting normal cells against oxidative damage and electrophilic attack.It is highly expressed in many solid tumors,suggesting a role in cancer development and progression.However,the role of NQO1 in gastric cancer and its effect on cancer development and prognosis have not been fully investigated.AIM To investigate the clinical relevance of NQO1 protein expression in gastric cancer and to explore the potential of NQO1 to serve as a prognostic biomarker and therapeutic target.METHODS In this retrospective study,gastric cancer specimens of 175 patients who were treated between 1995 and 2011 were subjected to immunohistochemistry analyses for NQO1.The correlation of NQO1 expression with gastric cancer prognosis and clinical and pathological parameters was investigated.RESULTS NQO1 protein was overexpressed in 59.43%(104/175)of the analyzed samples.Overexpression of NQO1 was associated with a significantly inferior prognosis.In addition,multivariate analysis suggested that NQO1 overexpression,along with tumor stage and patient age,are prominent prognostic biomarkers for gastric cancer.Moreover,NQO1 overexpression was correlated to a better response to 5-fluorouracil(5-FU)-based adjuvant chemotherapy.CONCLUSION NQO1 overexpression is associated with a significantly poor prognosis and better response to 5-FU in patients with gastric cancer.These findings are relevant for improving therapeutic approaches for gastric cancer patients.展开更多
AIM： To investigate the expression and activity of NAD（P）H quinone oxidoreductase 1 （NQO1） in human liver specimens obtained from patients with liver damage due to acetaminophen （APAP） overdose or primary bilia...AIM： To investigate the expression and activity of NAD（P）H quinone oxidoreductase 1 （NQO1） in human liver specimens obtained from patients with liver damage due to acetaminophen （APAP） overdose or primary biliary cirrhosis （PBC）. METHODS： NQOt activity was determined in cytosol from normal, APAP and PBC liver specimens. Western blot and immunohistochemical staining were used to determine patterns of NQO1 expression using a specific antibody against NQO1. RESULTS： NQO1 protein was very low in normal human livers. In both APAP and PBC livers, there was strong induction of NQO1 protein levels on Western blot. Correspondingly, significant up-regulation of enzyme activity （16- and 22-fold, P〈0.05） was also observed in APAP and PBC livers, respectively. Immunohistochemical analysis highlighted injury-specific patterns of NQO1 staining in both APAP and PBC livers. CONCLUSION： These data demonstrate that NQO1 protein and activity are markedly induced in human livers during both APAP overdose and PBC. Up-regulation of this cytoprotective enzyme may represent an adaptive stress response to limit further disease progression by detoxifying reactive species.展开更多
2-ketoisovalerate ferredoxin oxidoreductase (VOR) is a key enzyme in hyperthermophiles catalyzing the coenzyme A-dependent oxidative decarboxylation of aliphatic amino acid-derived 2-keto acids. The enzyme purified un...2-ketoisovalerate ferredoxin oxidoreductase (VOR) is a key enzyme in hyperthermophiles catalyzing the coenzyme A-dependent oxidative decarboxylation of aliphatic amino acid-derived 2-keto acids. The enzyme purified under anaerobic conditions from a hyperthermophilic archaeon, Thermococcus profundus, is a hetero-octamer (αβγδ)2 consisting of four different subunits, α = 45 kDa, β = 31 kDa, γ = 22 kDa and δ = 13 kDa, respectively, and it has three [4Fe-4S] clusters per αβγδ-protomer, similar to other ferredoxin oxidoreductases. In the present study, the native enzyme was purified from this strain and crystallized to give rod-like crystals that were suitable for X-ray diffraction experiments. The crystals belonged to space group P41212, with unit-cell parameters a = b = 136.20 ?, c = 221.07 ?. Diffraction images were processed to a resolution of 3.0 ?. The data collected so far indicate the approximate molecular boundaries and a partial main-chain trace of the enzyme.展开更多
The toxicity of acetaldehyde and age related changes on oxidoreductases in the liver,brain, kidney, and musele of female albino rats (Wistar strain) were studied. The specific activities of lactate [LDH], isocitrate [...The toxicity of acetaldehyde and age related changes on oxidoreductases in the liver,brain, kidney, and musele of female albino rats (Wistar strain) were studied. The specific activities of lactate [LDH], isocitrate [ICDH (NAD/NADP)], succinate [SDH], malate [MDH], glutamate [GDH] and glucose-6-Phosphate [G-6-PDH] dehydrogenases were signifieantly inereased as a function of age. However, acetaldehyde treatment significantly inhibited oxidoreductases in the tissues of 21, 90 and 180 day old rats. Liver enzymes of young (21 days) rats exhibited greater sensitivity to acetaldehyde toxicity. Similar inhibition of oxidoreductases in brain and kidney of adult (180 days) rats treated with acetaldehyde was observed. LDH and GDH as compared to other enzymes studied showed higher susceptibility to acetaldehyde toxicity. The differential sensitivity of tissues and inhibition of oxidoreductases by acetaldehyde as a function of age could be attributed to hypoxic conditions, energy crisis, and mitochondrial structural changes. The results suggest that acetaldehyde affects oxidation of glucose via HMP shuni pathway, glycolytic pathway and Krebs cycle resulting in the impairment of carbohydrate metabolism展开更多
Azoreductases are involved in the bioremediation by bacteria of azo dyes found in waste water.In the gut flora,they activate azo pro-drugs,which are used for treatment of inflammatory bowel disease,releasing the activ...Azoreductases are involved in the bioremediation by bacteria of azo dyes found in waste water.In the gut flora,they activate azo pro-drugs,which are used for treatment of inflammatory bowel disease,releasing the active component 5-aminosalycilic acid.The bacterium P.aeruginosa has three azoreductase genes,paAzoR1,paAzoR2 and paAzoR3,which as recombinant enzymes have been shown to have different substrate specificities.The mechanism of azoreduction relies upon tautomerisation of the substrate to the hydrazone form.We report here the characterization of the P.aeruginosa azoreductase enzymes,including determining their thermostability,cofactor preference and kinetic constants against a range of their favoured substrates.The expression levels of these enzymes during growth of P.aeruginosa are altered by the presence of azo substrates.It is shown that enzymes that were originally described as azoreductases,are likely to act as NADH quinone oxidoreductases.The low sequence identities observed among NAD(P)H quinone oxidoreductase and azoreductase enzymes suggests convergent evolution.展开更多
The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues.NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers.Therefore,it could be a ta...The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues.NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers.Therefore,it could be a target for cancer therapy.Because it is a quinone reductase,many bioimaging probes based on quinone structures target NAD(P)H:quinone oxidoreductase 1 to diagnose tumours.Its expression is higher in tumours than in normal tissues,and using target drugs such asβ-lapachone to reduce side effects in normal tissues can help.However,the physicochemical properties ofβlapachone limit its application.The problem can be solved by using nanosystems to deliverβ-lapachone.This minireview summarizes quinone-based fluorescent,nearinfrared and two-photon fluorescent probes,as well as nanosystems for delivering the NAD(P)H:quinone oxidoreductase 1-activating drugβ-lapachone.This review provides valuable information for the future development of probes and nano-delivery systems that target NAD(P)H:quinone oxidoreductase 1.展开更多
Ferredoxin-NADP＋ oxidoreductase （FNR）, functioning in the last step of the photosynthetic electron transfer chain, exists both as a soluble protein in the chloroplast stroma and tightly attached to chloroplast memb...Ferredoxin-NADP＋ oxidoreductase （FNR）, functioning in the last step of the photosynthetic electron transfer chain, exists both as a soluble protein in the chloroplast stroma and tightly attached to chloroplast membranes. Surface plasmon resonance assays showed that the two FNR isoforms, LFNR1 and LFNR2, are bound to the thylakoid membrane via the C-terminal domains of Tic62 and TROL proteins in a pH-dependent manner. The tic62 trol double mutants contained a reduced level of FNR, exclusively found in the soluble stroma. Although the mutant plants showed no visual phenotype or defects in the function of photosystems under any conditions studied, a low ratio of NADPH/NADP~ was detected. Since the CO2 fixation capacity did not differ between the tic62 trol plants and wild-type, it seems that the plants are able to funnel reducing power to most crucial reactions to ensure survival and fitness of the plants. However, the activity of malate dehydrogenase was down-regulated in the mutant plants. Apparently, the plastid metabolism is able to cope with substantial changes in directing the electrons from the light reactions to stromal metabolism and thus only few differences are visible in steady-state metabolite pool sizes of the tic62 trol plants.展开更多
Importance:Cytochrome P450 oxidoreductase deficiency (PORD) is a rare disease exhibiting a variety of clinical manifestations.This condition specifically leads to disordered steroidogenesis,which can affect the develo...Importance:Cytochrome P450 oxidoreductase deficiency (PORD) is a rare disease exhibiting a variety of clinical manifestations.This condition specifically leads to disordered steroidogenesis,which can affect the development of the reproductive system,skeleton,and other parts of the body.The severe form of PORD is difficult to differentiate with Antley-Bixler syndrome (ABS).The genetic characters and clinical evaluation of PORD are still unclear in China.Objective:To perform an exome analysis and identify the pathogenic cause in order to assist clinicians to obtain a proper evaluation on the genetic condition.Methods:The proband underwent detailed physical evaluations.DNA of the proband and his parents was isolated and whole-exome sequencing (WES) was performed.Variants were analyzed and evaluation according to the ACMG guideline.Results:A 1-year-old Chinese boy with midface hypoplasia,choanal stenosis,multiple joint contractures,micropenis and right cryptorchidism was misdiagnosed with Crouzon syndrome.By trio-whole-exome sequencing,we identified an unreported compound heterozygous mutation (c.667C＞T,p.R223* and c.1370G＞A,p.R457H) in POR in the proband.This mutation was inherited from healthy heterozygous parents,supporting the diagnosis of PORD,which was further confirmed by biochemical characteristics.Interpretation:We have identified a pathogenic variant with an unreported compound heterozygous POR mutation,which expands the clinical and genetic spectra of PORD and emphasizes the usefulness of WES for genetic diagnosis.展开更多
A membrane-bound protein was purified from rat liver mitochondria.After being di-gested with V8 protease,two peptides containing identical 14 amino acid residue sequences were obtained.Using the 14 amino acid peptide ...A membrane-bound protein was purified from rat liver mitochondria.After being di-gested with V8 protease,two peptides containing identical 14 amino acid residue sequences were obtained.Using the 14 amino acid peptide derived DNA sequence as gene specific primer,the cDNA of correspondent gene 5′-terminal and 3′-terminal were obtained by RACE technique.The full-length cDNA that encoded a protein of 616 amino acids was thus cloned,which included the above mentioned peptide sequence.The full length cDNA was highly homologous to that of human ETF-QO,indicating that it may be the cDNA of rat ETF-QO.ETF-QO is an iron sulfur protein located in mitochondria inner membrane containing two kinds of redox center:FAD and[4Fe-4S]center.After comparing the sequence from the cDNA of the 616 amino acids protein with that of the mature protein of rat liver mitochondria,it was found that the N terminal 32 amino acid residues did not exist in the mature protein,indicating that the cDNA was that of ETF-QOp.When the cDNA was expressed in Saccharomyces cerevisiae with inducible vectors,the protein product was enriched in mitochondrial fraction and exhibited electron transfer activity(NBT re-ductase activity)of ETF-QO.Results demonstrated that the 32 amino acid peptide was a mito-chondrial targeting peptide,and both FAD and iron-sulfur cluster were inserted properly into the expressed ETF-QO.ETF-QO had a high level expression in rat heart,liver and kidney.The fu-sion protein of GFP-ETF-QO co-localized with mitochondria in COS-7 cells.展开更多
Acidithiobacillus caldus,a typical sulfur oxidizer,derives the majority of its energy from sulfur oxidation.And the essential enzyme for sulfide oxidation catalysis is sulfide quinone oxidoreductase(SQR),an ancient fl...Acidithiobacillus caldus,a typical sulfur oxidizer,derives the majority of its energy from sulfur oxidation.And the essential enzyme for sulfide oxidation catalysis is sulfide quinone oxidoreductase(SQR),an ancient flavoprotein.Here,the catalytic mechanism of SQR generated from A.caldus was investigated(SQR^(Ac)).According to phylogenetic study,SQR^(Ac)(ACAty RS11315)is closely related to SQR(BAD99305)of Acidithiobacillus ferrooxidans NASF-1 and is classified as a type I Sqr enzyme.SQR^(Ac)heterologously produced in Escherichia coli exhibits the distinctive absorption peaks(375,450 nm)of the flavoproteins family of proteins in its absorption spectrum.Utilizing site-directed mutagenesis,the function of conserved cysteines in the catalytic pathway was determined.Based on the sulfide quinone redox reactions in vitro of SQR^(Ac)and variations,Cys160 and Cys356 have been identified as enzyme-active residues.Mutation of another cysteine present in all type I SQRs(Cys128)decreased enzyme activity by 56%,indicating that this residue plays an important but non-essential role in enzyme function.In addition,the binding affinities of SQR^(Ac),the visualization of its 3D structure,and the interaction between receptors and ligands were investigated.Finally,a suitable sulfide quinone redox catalytic mechanism for A.caldus was proposed.展开更多
Ferroptosis,a type of programmed cell death,represents a distinct paradigm in cell biology.It is characterized by the iron-dependent accumulation of reactive oxygen species,which induce lipid peroxidation(LPO),and is ...Ferroptosis,a type of programmed cell death,represents a distinct paradigm in cell biology.It is characterized by the iron-dependent accumulation of reactive oxygen species,which induce lipid peroxidation(LPO),and is orchestrated by the interplay between iron,lipid peroxides,and glutathione.In this review,we emphasize the frequently overlooked role of iron in LPO beyond the classical iron-driven Fenton reaction in several crucial processes that regulate cellular iron homeostasis,including iron intake and export as well as ferritinophagy,and the emerging roles of endoplasmic reticulum-resident flavoprotein oxidoreductases,especially P450 oxidoreductases,in modulating LPO.We summarize how various types of fatty acids(FAs),including saturated,monounsaturated,and polyunsaturated FAs,differentially influence ferroptosis when incorporated into phospholipids.Furthermore,we highlight the therapeutic potential of targeting LPO to mitigate ferroptosis and discuss the regulatory mechanisms of endogenous lipophilic radical-trapping antioxidants that confer resistance to ferroptosis,shedding light on therapeutic avenues for ferroptosis-associated diseases.展开更多
Biocatalysis with nicotinamide adenine dinucleotide phosphate(NADP)-dependent oxidoreductases faces a challenge in improving the efficiency of the costly cofactor utilization.Although enzyme fusion can offer cofactor ...Biocatalysis with nicotinamide adenine dinucleotide phosphate(NADP)-dependent oxidoreductases faces a challenge in improving the efficiency of the costly cofactor utilization.Although enzyme fusion can offer cofactor regeneration,the high-volume input and limited cofactor recyclability still make the enzymatic processes unsustainable.Therefore,it is of great significance to reduce cofactor input in a fusion enzyme(FuE)system,but no successful practice has been reported.Herein,we design a decapeptide bridge,RRRQRRRARR(R10),with high affinity for NADPH to construct fusion oxidoreductases(phenylacetone monooxygenase and phosphite dehydrogenase)for ester synthesis and NADP recycling.The peptide bridge enables electrostatic cofactor channeling that transports NADPH/NADP^(+)across the peptide between the enzymes’NADP-binding pockets,so the fusion enzyme(FuE-R10)presents 2.1-folds and 2.0-folds higher conversions than mixed free enzymes and a flexible linker(GGGGSGGGGS)-fused enzyme,respectively,at NADPH/FuE of 0.1.The fusion enzyme,FuE-R5,bridged by a half-shortened linker,is proved more effective in facilitating cofactor channeling;compared to the mixed free enzymes,FuE-R5 exhibits two orders of magnitude reduction of NADPH input in ester synthesis.The work has thus demonstrated the potential of the cofactor bridging strategy in the development of sustainable cofactor-dependent cascade biocatalysis.展开更多
Angiosperms need light to synthesize chlorophyll, but lotus (Nelumbo nucifera Gaertn.) embryo was suspected to have the ability to form chlorophyll in the dark because lotus embryo can turn into green under the covera...Angiosperms need light to synthesize chlorophyll, but lotus (Nelumbo nucifera Gaertn.) embryo was suspected to have the ability to form chlorophyll in the dark because lotus embryo can turn into green under the coverage of four layers of integuments (cotyledon, seed coat, pericarp, lotus pod) which were thought impossible for light to pass through. The authors excluded this possibility based on two experimental results: First, enclosing the young lotus pod with aluminium foil, the growth of louts embryo continued, but the chlorophyll formation was seriously inhibited. A lot of protochlorophyllide, chlorophyll precursor, were accumulated, most of which were combined with LPOR (light dependent protochlorophyllide oxidoreductase). Second, DPOR (dark or light-independent protochlorophyllide oxidoreductase) was the enzyme necessary for chlorophyll synthesis in the dark. The genes encoding DPOR were conservative in many species, but no homologues could be found in lotus genome. Taken together, authers' results clearly demonstrated that lotus embryo synthesizes chlorophyll only through the light-dependent pathway.展开更多
CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzy...CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzymes for CO2 fixation in response to salinity, pH and O2 in Medium 829 were also investigated. The results demonstrate that C. profundus contained aclB, porA and oorA, the genes encoding key enzymes of reductive tricarboxylic acid （rTCA） cycle. However, genes fragments of cbbL and cbbMencoding key enzyme of Calvin cycle were not recovered. Key enzymatic activities of ATP citrate lyase （ACL）, pyruvate： ferredoxin oxidoreductase （POR） and 2-oxoglutarate： ferredoxin oxidoreductase （OOR） were also present in C. profun- dus. The combination of genetic and enzymatic analyses confirm that C. profundus adopted rTCA cycle for carbon assimilation. The results of aclB and oorA relative expressions of C. profundus demonstrate that the ranges of environmental factors for high genes expression were sea salt 3.0%-5.0% （optimum 3.0%）, pH 5.0-6.5（optimum pH 6.5）, anaerobic to microaerobic conditions （optimum 1.0% 02）. Gene expression pat- terns under different conditions show similar patterns with bacterial growth, revealing that key rTCA cycle genes provided molecular basis for bacterial growth and propagation. Our results suggest that C. profun- dus could regulate key genes of rTCA cycle for carbon assimilation and energy metabolism in response to environmental fluctuations in hydrothermal vent.展开更多
Background:Aldo-keto oxidoreductase(AKR)inhibitors could reverse the resistance of several cancer cells to cis-platin,but their role in resistance remains unclear.Methods:We verified the difference of AKR1Cs expressio...Background:Aldo-keto oxidoreductase(AKR)inhibitors could reverse the resistance of several cancer cells to cis-platin,but their role in resistance remains unclear.Methods:We verified the difference of AKR1Cs expression by Western blot,RNA sequencing and qRT-PCR.The differences of AKR1Cs expression were analyzed and inferred.Use Assay of NADH and NAD^(+)content to verify the inference.The Docking experience was used to verify the affinity between MPA,MCFLA,MLS and AKR1C3.Results:Our RNA-seq results showed de novo NAD biosynthesis-related genes and NAD(P)H-dependent oxidoreductases were significantly upregulated in cis-platin-resistant HepG2 hepatic cancer cells(HepG2-RC cells)compared with HepG2 cells.At least 63 NAD(P)H-dependent reductase/oxidases were upregulated in HepG2-RC cells at least twofold.Knockdown of AKR1Cs could increase cis-platin sensitivity in HepG2-RC cells about two-fold.Interestingly,the AKR1C inhibitor meclofenamic acid could increase the cis-platin sensitivity of HepG2-RC cells about eight-fold,indicating that the knockdown of AKR1Cs only partially reversed the resistance.Meanwhile,the amount of total NAD and the ratio of NADH/NAD^(+)were increased in HepG2-RC cells compared with HepG2 cells.The ratio of NADH/NAD^(+)in HepG2-RC cells was almost seven-fold higher than in HepG2 or HL-7702 cells.Increased NADH expression could be explained as a directly operating antioxidant to scavenge cis-platin-induced radicals.Conclusion:We report here that NADH,which is produced by NAD(P)Hdependent oxidoreductases,plays a key role in the AKR-associated cis-platin resistance of HepG2 hepatic cancer cells.展开更多
AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2(Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group(A);CCl...AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2(Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group(A);CCl4-induced hepatic fibrosis group(B);blueberry prevention group(C);Dan-shao-hua-xian capsule(DSHX) prevention group(D);and blueberry + DSHX prevention group(E).Liver fibrosis was induced in rats by subcutaneous injection of CCl4 and a high-lipid/low-protein diet for 8 wk(except the control group).The level of hyaluronic acid(HA) and alanine aminotransferase(ALT) in serum was examined.The activity of superoxide dismutase(SOD),glutathione-S-transferase(GST) and malondialdehyde(MDA) in liver homogenates was determined.The degree of hepatic fibrosis was evaluated by hematoxylin and eosin and Masson staining.Expression of Nrf2 and NADPH quinone oxidoreductase 1(Nqo1) was detected by real-time reversed transcribed-polymerase chain reaction,immunohistochemical techniques,and western blotting.RESULTS:Compared with group B,liver indices,levels of serum HA and ALT of groups C,D and E were reduced(liver indices:0.038 ± 0.008,0.036 ± 0.007,0.036 ± 0.005 vs 0.054 ± 0.009,P<0.05;HA:502.33 ± 110.57 ng/mL,524.25 ± 255.42 ng/mL,499.25 ± 198.10 ng/mL vs 828.50 ± 237.83 ng/mL,P<0.05;ALT:149.44 ± 16.51 U/L,136.88 ± 10.07 U/L,127.38 ± 11.03 U/L vs 203.25 ± 31.62 U/L,P<0.05),and SOD level was significantly higher,but MDA level was lower,in liver homogenates(SOD:1.36 ± 0.09 U/mg,1.42 ± 0.13 U/mg,1.50 ± 0.15 U/mg vs 1.08 ± 0.19 U/mg,P<0.05;MDA:0.294 ± 0.026 nmol/mg,0.285 ± 0.025 nmol/mg,0.284 ± 0.028 nmol/mg vs 0.335 ± 0.056 nmol/mg,P<0.05).Meanwhile,the stage of hepatic fibrosis was significantly weakened(P<0.05).Compared with group A,the activity of GST liver homogenates and expression levels of Nrf2 and Nqo1 in group B were elevated(P<0.05).The expression level of Nrf2 and Nqo1 in groups C,D,and E were increased as compared with group B,but the difference was not significant.CONCLUSION:Blueberry has preventive and protective effects on CCl4-induced hepatic fibrosis by reducing hepatocyte injury and lipid peroxidation.However,these effects may not be related to the activation of Nrf2 during long-term of CCl4.展开更多
No study has been performed on identifying microRNAs(miRNAs) and their targets in cotton although cotton is one of the most important fiber and economic crops around the world.In this study,
In order to improve beef color and color stability, step-chilling （SC） was applied on excised bovine Iongissimus lumborum muscle, with chilling starting at 0-4℃ for 5 h, then holding the temperature at 12-18℃ for ...In order to improve beef color and color stability, step-chilling （SC） was applied on excised bovine Iongissimus lumborum muscle, with chilling starting at 0-4℃ for 5 h, then holding the temperature at 12-18℃ for 6 h, followed by 0-4℃ again until 24 h post-mortem, pH and temperature were measured during rigor on SC loins as well as those subjected to routine chilling （RC, 0-4℃, till 24 h post-mortem）. Color L＊, a＊, b＊ values, metmyoglobin （MetMb） content, MetMb reducing ability （MRA） and NADH content were determined on samples aged for 1, 7, and 14 d. Sarcoplasmic proteome analysis was only conducted on d 1 samples. The results showed muscles subjected to SC maintained a temperature at around 15℃ for 5 to 10 h post-mortem, and exhibited a slow temperature decline, but rapid pH decline. Beef steaks treated with SC had higher L＊, a＊, b＊ and chroma values than those of RC samples at 1 and 7 d chilled storage （0-4℃）, while showing no significant difference for a＊, b＊ and chroma values at d 14. The SC samples also exhibited a lower relative content of surface MetMb, higher MRA and NADH content, compared with RC beef steaks during storage, indicating the SC-treated beef showed an improved color stability. Eleven differential protein spots/nine proteins were identified by two-dimensional gel electrophoresis and mass spectrometry, and those proteins were mainly involved in redox, chaperone binding, metabolic and peroxidase activity. Oxidoreductases play a role in decreasing the oxidation-induced myoglobin oxidation and benefiting the production of NADH, and finally improving the colour of beef. Of these, pyruvate dehydrogenase E1 component subunit beta showed a positive correlation with color L＊, a＊, b＊ values and accounted for more than 60% of the variation in color values; this protein can be considered as a potential beef color biomarker. The present study provided valuable information for studies on the molecular mechanism of color improvement from step-chilling, as well as for identifying markers associated with beef color.展开更多
The WW domain-containing oxidoreductase（WWOX） is a tumor suppressor in a variety of cancers, including breast cancer. Reduced WWOX expression is associated with the basal-like subtype and a relatively poor disease-f...The WW domain-containing oxidoreductase（WWOX） is a tumor suppressor in a variety of cancers, including breast cancer. Reduced WWOX expression is associated with the basal-like subtype and a relatively poor disease-free survival rate among breast cancer patients. Though several WWOX partners have been identified, the functional mechanisms of WWOX＇s role in cancers have not been fully addressed to date. In the current study, we found WWOX suppresses expression of KLF5—an oncogenic transcription factor—at protein level, and suppresses cancer cell proliferation in both bladder and breast cancer cell lines. Furthermore, we demonstrated that WWOX physically interacts with KLF5 via the former＇s WW domains and the latter＇s PY motifs. Interestingly, we found the expression of WWOX negatively correlates with KLF5 expression in a panel of breast cancer cell lines. Taken together, we conjecture that WWOX may suppress cancer cell proliferation partially by reducing the expression of KLF5.展开更多
Glyphosate is an important organophosphonate herbicide used to eliminate grasses and herbaceous plants in many vegetation management situations.Its extensive use is causing environmental pollution,and consequently,the...Glyphosate is an important organophosphonate herbicide used to eliminate grasses and herbaceous plants in many vegetation management situations.Its extensive use is causing environmental pollution,and consequently,there is a need to remove it from the environment using an eco-friendly and cost-effective method.As a step to address this problem,a novel bacterial strain Comamonas odontotermitis F2,capable to utilize glyphosate as a carbon(C)and/or phosphorus(P)source,was isolated from a glyphostate-contaminated field soil in Australia and characterized.Response surface methodology(RSM)employing a 2^3 full factorial central composite design was used to optimize glyphosate degradation by C.odontotermitis P2 under various culture conditions.The strain C.odontotermitis P2 was proficient in degrading 1.5 g L^-1 glyphosate completely within 104 h.The optimal conditions for the degradation of glyphosate were found to be pH 7.4,29.9℃,and an inoculum density of 0.54 g L^-1,resulting in a maximum degradation of 90%.Sequencing of glyphosate oxidoreductase(GOX)and C-P lyase(phnJ)genes from C.odontotermitis P2 revealed 999c and 93%identities to already reported bacterial GOX and phnJ genes,respectively.The presence of these two genes in C.odontotermitis indicates its potential to degrade glyphosate through GOX and C-P lyase metabolic pathways.This study demonstrates the potential of C.odontotermitis P2 for efficient degradation of glyphosate,which can be exploited for remediation of glyphosate.展开更多
基金Supported by the National Natural Science Foundation of China,No.31971188 and No.81773189the Nature Science Foundation of Zhejiang Province,China,No.LQ16H160004 and No.LY17H270002The Hygiene Department of Zhejiang,No.2016KYB139.
文摘BACKGROUND Quinine oxidoreductase 1(NQO1)plays a vital role in protecting normal cells against oxidative damage and electrophilic attack.It is highly expressed in many solid tumors,suggesting a role in cancer development and progression.However,the role of NQO1 in gastric cancer and its effect on cancer development and prognosis have not been fully investigated.AIM To investigate the clinical relevance of NQO1 protein expression in gastric cancer and to explore the potential of NQO1 to serve as a prognostic biomarker and therapeutic target.METHODS In this retrospective study,gastric cancer specimens of 175 patients who were treated between 1995 and 2011 were subjected to immunohistochemistry analyses for NQO1.The correlation of NQO1 expression with gastric cancer prognosis and clinical and pathological parameters was investigated.RESULTS NQO1 protein was overexpressed in 59.43%(104/175)of the analyzed samples.Overexpression of NQO1 was associated with a significantly inferior prognosis.In addition,multivariate analysis suggested that NQO1 overexpression,along with tumor stage and patient age,are prominent prognostic biomarkers for gastric cancer.Moreover,NQO1 overexpression was correlated to a better response to 5-fluorouracil(5-FU)-based adjuvant chemotherapy.CONCLUSION NQO1 overexpression is associated with a significantly poor prognosis and better response to 5-FU in patients with gastric cancer.These findings are relevant for improving therapeutic approaches for gastric cancer patients.
文摘AIM： To investigate the expression and activity of NAD（P）H quinone oxidoreductase 1 （NQO1） in human liver specimens obtained from patients with liver damage due to acetaminophen （APAP） overdose or primary biliary cirrhosis （PBC）. METHODS： NQOt activity was determined in cytosol from normal, APAP and PBC liver specimens. Western blot and immunohistochemical staining were used to determine patterns of NQO1 expression using a specific antibody against NQO1. RESULTS： NQO1 protein was very low in normal human livers. In both APAP and PBC livers, there was strong induction of NQO1 protein levels on Western blot. Correspondingly, significant up-regulation of enzyme activity （16- and 22-fold, P〈0.05） was also observed in APAP and PBC livers, respectively. Immunohistochemical analysis highlighted injury-specific patterns of NQO1 staining in both APAP and PBC livers. CONCLUSION： These data demonstrate that NQO1 protein and activity are markedly induced in human livers during both APAP overdose and PBC. Up-regulation of this cytoprotective enzyme may represent an adaptive stress response to limit further disease progression by detoxifying reactive species.
文摘2-ketoisovalerate ferredoxin oxidoreductase (VOR) is a key enzyme in hyperthermophiles catalyzing the coenzyme A-dependent oxidative decarboxylation of aliphatic amino acid-derived 2-keto acids. The enzyme purified under anaerobic conditions from a hyperthermophilic archaeon, Thermococcus profundus, is a hetero-octamer (αβγδ)2 consisting of four different subunits, α = 45 kDa, β = 31 kDa, γ = 22 kDa and δ = 13 kDa, respectively, and it has three [4Fe-4S] clusters per αβγδ-protomer, similar to other ferredoxin oxidoreductases. In the present study, the native enzyme was purified from this strain and crystallized to give rod-like crystals that were suitable for X-ray diffraction experiments. The crystals belonged to space group P41212, with unit-cell parameters a = b = 136.20 ?, c = 221.07 ?. Diffraction images were processed to a resolution of 3.0 ?. The data collected so far indicate the approximate molecular boundaries and a partial main-chain trace of the enzyme.
文摘The toxicity of acetaldehyde and age related changes on oxidoreductases in the liver,brain, kidney, and musele of female albino rats (Wistar strain) were studied. The specific activities of lactate [LDH], isocitrate [ICDH (NAD/NADP)], succinate [SDH], malate [MDH], glutamate [GDH] and glucose-6-Phosphate [G-6-PDH] dehydrogenases were signifieantly inereased as a function of age. However, acetaldehyde treatment significantly inhibited oxidoreductases in the tissues of 21, 90 and 180 day old rats. Liver enzymes of young (21 days) rats exhibited greater sensitivity to acetaldehyde toxicity. Similar inhibition of oxidoreductases in brain and kidney of adult (180 days) rats treated with acetaldehyde was observed. LDH and GDH as compared to other enzymes studied showed higher susceptibility to acetaldehyde toxicity. The differential sensitivity of tissues and inhibition of oxidoreductases by acetaldehyde as a function of age could be attributed to hypoxic conditions, energy crisis, and mitochondrial structural changes. The results suggest that acetaldehyde affects oxidation of glucose via HMP shuni pathway, glycolytic pathway and Krebs cycle resulting in the impairment of carbohydrate metabolism
文摘Azoreductases are involved in the bioremediation by bacteria of azo dyes found in waste water.In the gut flora,they activate azo pro-drugs,which are used for treatment of inflammatory bowel disease,releasing the active component 5-aminosalycilic acid.The bacterium P.aeruginosa has three azoreductase genes,paAzoR1,paAzoR2 and paAzoR3,which as recombinant enzymes have been shown to have different substrate specificities.The mechanism of azoreduction relies upon tautomerisation of the substrate to the hydrazone form.We report here the characterization of the P.aeruginosa azoreductase enzymes,including determining their thermostability,cofactor preference and kinetic constants against a range of their favoured substrates.The expression levels of these enzymes during growth of P.aeruginosa are altered by the presence of azo substrates.It is shown that enzymes that were originally described as azoreductases,are likely to act as NADH quinone oxidoreductases.The low sequence identities observed among NAD(P)H quinone oxidoreductase and azoreductase enzymes suggests convergent evolution.
基金the financial support from the Tianjin Science and Technology Committee(Grant No.19JCYBJC28400)the Basic Research General Program of Shenzhen Science and Technology Innovation Commission in 2020(Grant No.JCYJ20190806162412752).
文摘The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues.NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers.Therefore,it could be a target for cancer therapy.Because it is a quinone reductase,many bioimaging probes based on quinone structures target NAD(P)H:quinone oxidoreductase 1 to diagnose tumours.Its expression is higher in tumours than in normal tissues,and using target drugs such asβ-lapachone to reduce side effects in normal tissues can help.However,the physicochemical properties ofβlapachone limit its application.The problem can be solved by using nanosystems to deliverβ-lapachone.This minireview summarizes quinone-based fluorescent,nearinfrared and two-photon fluorescent probes,as well as nanosystems for delivering the NAD(P)H:quinone oxidoreductase 1-activating drugβ-lapachone.This review provides valuable information for the future development of probes and nano-delivery systems that target NAD(P)H:quinone oxidoreductase 1.
文摘Ferredoxin-NADP＋ oxidoreductase （FNR）, functioning in the last step of the photosynthetic electron transfer chain, exists both as a soluble protein in the chloroplast stroma and tightly attached to chloroplast membranes. Surface plasmon resonance assays showed that the two FNR isoforms, LFNR1 and LFNR2, are bound to the thylakoid membrane via the C-terminal domains of Tic62 and TROL proteins in a pH-dependent manner. The tic62 trol double mutants contained a reduced level of FNR, exclusively found in the soluble stroma. Although the mutant plants showed no visual phenotype or defects in the function of photosystems under any conditions studied, a low ratio of NADPH/NADP~ was detected. Since the CO2 fixation capacity did not differ between the tic62 trol plants and wild-type, it seems that the plants are able to funnel reducing power to most crucial reactions to ensure survival and fitness of the plants. However, the activity of malate dehydrogenase was down-regulated in the mutant plants. Apparently, the plastid metabolism is able to cope with substantial changes in directing the electrons from the light reactions to stromal metabolism and thus only few differences are visible in steady-state metabolite pool sizes of the tic62 trol plants.
基金National Natural Science Foundation of China(81670789,31401171)%Ministry of Science and Technology of China(2016YFC1000306)%The Beijing Municipal Science and Technology Commission Foundation(Z141100002114009)%Beijing Municipal Commission of Health and Family Planning Foundation(PXM2017_026274_000001)%Beijing Municipal Administration of Hospitals Foundation(QML20161201)
文摘Importance:Cytochrome P450 oxidoreductase deficiency (PORD) is a rare disease exhibiting a variety of clinical manifestations.This condition specifically leads to disordered steroidogenesis,which can affect the development of the reproductive system,skeleton,and other parts of the body.The severe form of PORD is difficult to differentiate with Antley-Bixler syndrome (ABS).The genetic characters and clinical evaluation of PORD are still unclear in China.Objective:To perform an exome analysis and identify the pathogenic cause in order to assist clinicians to obtain a proper evaluation on the genetic condition.Methods:The proband underwent detailed physical evaluations.DNA of the proband and his parents was isolated and whole-exome sequencing (WES) was performed.Variants were analyzed and evaluation according to the ACMG guideline.Results:A 1-year-old Chinese boy with midface hypoplasia,choanal stenosis,multiple joint contractures,micropenis and right cryptorchidism was misdiagnosed with Crouzon syndrome.By trio-whole-exome sequencing,we identified an unreported compound heterozygous mutation (c.667C＞T,p.R223* and c.1370G＞A,p.R457H) in POR in the proband.This mutation was inherited from healthy heterozygous parents,supporting the diagnosis of PORD,which was further confirmed by biochemical characteristics.Interpretation:We have identified a pathogenic variant with an unreported compound heterozygous POR mutation,which expands the clinical and genetic spectra of PORD and emphasizes the usefulness of WES for genetic diagnosis.
基金This project was supported by the Knowledge Innovation Program of the Chinese Academy of Sciences.
文摘A membrane-bound protein was purified from rat liver mitochondria.After being di-gested with V8 protease,two peptides containing identical 14 amino acid residue sequences were obtained.Using the 14 amino acid peptide derived DNA sequence as gene specific primer,the cDNA of correspondent gene 5′-terminal and 3′-terminal were obtained by RACE technique.The full-length cDNA that encoded a protein of 616 amino acids was thus cloned,which included the above mentioned peptide sequence.The full length cDNA was highly homologous to that of human ETF-QO,indicating that it may be the cDNA of rat ETF-QO.ETF-QO is an iron sulfur protein located in mitochondria inner membrane containing two kinds of redox center:FAD and[4Fe-4S]center.After comparing the sequence from the cDNA of the 616 amino acids protein with that of the mature protein of rat liver mitochondria,it was found that the N terminal 32 amino acid residues did not exist in the mature protein,indicating that the cDNA was that of ETF-QOp.When the cDNA was expressed in Saccharomyces cerevisiae with inducible vectors,the protein product was enriched in mitochondrial fraction and exhibited electron transfer activity(NBT re-ductase activity)of ETF-QO.Results demonstrated that the 32 amino acid peptide was a mito-chondrial targeting peptide,and both FAD and iron-sulfur cluster were inserted properly into the expressed ETF-QO.ETF-QO had a high level expression in rat heart,liver and kidney.The fu-sion protein of GFP-ETF-QO co-localized with mitochondria in COS-7 cells.
基金the National Key Research and Development Program of China(2022YFC3401300)the National Natural Science Foundation of China(No.21878128,21,776,113,31,701,582+2 种基金21,606,110)the Fundamental Research Funds for the Central Universities(No.2050205)Program of Introducing Talents of Discipline to Universities(No.111-2-06).
文摘Acidithiobacillus caldus,a typical sulfur oxidizer,derives the majority of its energy from sulfur oxidation.And the essential enzyme for sulfide oxidation catalysis is sulfide quinone oxidoreductase(SQR),an ancient flavoprotein.Here,the catalytic mechanism of SQR generated from A.caldus was investigated(SQR^(Ac)).According to phylogenetic study,SQR^(Ac)(ACAty RS11315)is closely related to SQR(BAD99305)of Acidithiobacillus ferrooxidans NASF-1 and is classified as a type I Sqr enzyme.SQR^(Ac)heterologously produced in Escherichia coli exhibits the distinctive absorption peaks(375,450 nm)of the flavoproteins family of proteins in its absorption spectrum.Utilizing site-directed mutagenesis,the function of conserved cysteines in the catalytic pathway was determined.Based on the sulfide quinone redox reactions in vitro of SQR^(Ac)and variations,Cys160 and Cys356 have been identified as enzyme-active residues.Mutation of another cysteine present in all type I SQRs(Cys128)decreased enzyme activity by 56%,indicating that this residue plays an important but non-essential role in enzyme function.In addition,the binding affinities of SQR^(Ac),the visualization of its 3D structure,and the interaction between receptors and ligands were investigated.Finally,a suitable sulfide quinone redox catalytic mechanism for A.caldus was proposed.
基金supported by grants from the National Natural Science Foundation of China(22076104)the“Taishan Scholars”Program for Young Expert of Shandong Province(tsqn202103105).
文摘Ferroptosis,a type of programmed cell death,represents a distinct paradigm in cell biology.It is characterized by the iron-dependent accumulation of reactive oxygen species,which induce lipid peroxidation(LPO),and is orchestrated by the interplay between iron,lipid peroxides,and glutathione.In this review,we emphasize the frequently overlooked role of iron in LPO beyond the classical iron-driven Fenton reaction in several crucial processes that regulate cellular iron homeostasis,including iron intake and export as well as ferritinophagy,and the emerging roles of endoplasmic reticulum-resident flavoprotein oxidoreductases,especially P450 oxidoreductases,in modulating LPO.We summarize how various types of fatty acids(FAs),including saturated,monounsaturated,and polyunsaturated FAs,differentially influence ferroptosis when incorporated into phospholipids.Furthermore,we highlight the therapeutic potential of targeting LPO to mitigate ferroptosis and discuss the regulatory mechanisms of endogenous lipophilic radical-trapping antioxidants that confer resistance to ferroptosis,shedding light on therapeutic avenues for ferroptosis-associated diseases.
文摘Biocatalysis with nicotinamide adenine dinucleotide phosphate(NADP)-dependent oxidoreductases faces a challenge in improving the efficiency of the costly cofactor utilization.Although enzyme fusion can offer cofactor regeneration,the high-volume input and limited cofactor recyclability still make the enzymatic processes unsustainable.Therefore,it is of great significance to reduce cofactor input in a fusion enzyme(FuE)system,but no successful practice has been reported.Herein,we design a decapeptide bridge,RRRQRRRARR(R10),with high affinity for NADPH to construct fusion oxidoreductases(phenylacetone monooxygenase and phosphite dehydrogenase)for ester synthesis and NADP recycling.The peptide bridge enables electrostatic cofactor channeling that transports NADPH/NADP^(+)across the peptide between the enzymes’NADP-binding pockets,so the fusion enzyme(FuE-R10)presents 2.1-folds and 2.0-folds higher conversions than mixed free enzymes and a flexible linker(GGGGSGGGGS)-fused enzyme,respectively,at NADPH/FuE of 0.1.The fusion enzyme,FuE-R5,bridged by a half-shortened linker,is proved more effective in facilitating cofactor channeling;compared to the mixed free enzymes,FuE-R5 exhibits two orders of magnitude reduction of NADPH input in ester synthesis.The work has thus demonstrated the potential of the cofactor bridging strategy in the development of sustainable cofactor-dependent cascade biocatalysis.
文摘Angiosperms need light to synthesize chlorophyll, but lotus (Nelumbo nucifera Gaertn.) embryo was suspected to have the ability to form chlorophyll in the dark because lotus embryo can turn into green under the coverage of four layers of integuments (cotyledon, seed coat, pericarp, lotus pod) which were thought impossible for light to pass through. The authors excluded this possibility based on two experimental results: First, enclosing the young lotus pod with aluminium foil, the growth of louts embryo continued, but the chlorophyll formation was seriously inhibited. A lot of protochlorophyllide, chlorophyll precursor, were accumulated, most of which were combined with LPOR (light dependent protochlorophyllide oxidoreductase). Second, DPOR (dark or light-independent protochlorophyllide oxidoreductase) was the enzyme necessary for chlorophyll synthesis in the dark. The genes encoding DPOR were conservative in many species, but no homologues could be found in lotus genome. Taken together, authers' results clearly demonstrated that lotus embryo synthesizes chlorophyll only through the light-dependent pathway.
基金China Ocean Mineral Resources R&D Association under contract No. DYXM-115-02-2-19Basic Scientific Fund for National Public Research Institutes of China under contract No. 2011T04
文摘CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzymes for CO2 fixation in response to salinity, pH and O2 in Medium 829 were also investigated. The results demonstrate that C. profundus contained aclB, porA and oorA, the genes encoding key enzymes of reductive tricarboxylic acid （rTCA） cycle. However, genes fragments of cbbL and cbbMencoding key enzyme of Calvin cycle were not recovered. Key enzymatic activities of ATP citrate lyase （ACL）, pyruvate： ferredoxin oxidoreductase （POR） and 2-oxoglutarate： ferredoxin oxidoreductase （OOR） were also present in C. profun- dus. The combination of genetic and enzymatic analyses confirm that C. profundus adopted rTCA cycle for carbon assimilation. The results of aclB and oorA relative expressions of C. profundus demonstrate that the ranges of environmental factors for high genes expression were sea salt 3.0%-5.0% （optimum 3.0%）, pH 5.0-6.5（optimum pH 6.5）, anaerobic to microaerobic conditions （optimum 1.0% 02）. Gene expression pat- terns under different conditions show similar patterns with bacterial growth, revealing that key rTCA cycle genes provided molecular basis for bacterial growth and propagation. Our results suggest that C. profun- dus could regulate key genes of rTCA cycle for carbon assimilation and energy metabolism in response to environmental fluctuations in hydrothermal vent.
基金supported by the Science and Technology Development Plan Project of Jilin Province,China[20200708101YY]The Foundation of Jilin Province Science and Technology Department[20200801062GH].
文摘Background:Aldo-keto oxidoreductase(AKR)inhibitors could reverse the resistance of several cancer cells to cis-platin,but their role in resistance remains unclear.Methods:We verified the difference of AKR1Cs expression by Western blot,RNA sequencing and qRT-PCR.The differences of AKR1Cs expression were analyzed and inferred.Use Assay of NADH and NAD^(+)content to verify the inference.The Docking experience was used to verify the affinity between MPA,MCFLA,MLS and AKR1C3.Results:Our RNA-seq results showed de novo NAD biosynthesis-related genes and NAD(P)H-dependent oxidoreductases were significantly upregulated in cis-platin-resistant HepG2 hepatic cancer cells(HepG2-RC cells)compared with HepG2 cells.At least 63 NAD(P)H-dependent reductase/oxidases were upregulated in HepG2-RC cells at least twofold.Knockdown of AKR1Cs could increase cis-platin sensitivity in HepG2-RC cells about two-fold.Interestingly,the AKR1C inhibitor meclofenamic acid could increase the cis-platin sensitivity of HepG2-RC cells about eight-fold,indicating that the knockdown of AKR1Cs only partially reversed the resistance.Meanwhile,the amount of total NAD and the ratio of NADH/NAD^(+)were increased in HepG2-RC cells compared with HepG2 cells.The ratio of NADH/NAD^(+)in HepG2-RC cells was almost seven-fold higher than in HepG2 or HL-7702 cells.Increased NADH expression could be explained as a directly operating antioxidant to scavenge cis-platin-induced radicals.Conclusion:We report here that NADH,which is produced by NAD(P)Hdependent oxidoreductases,plays a key role in the AKR-associated cis-platin resistance of HepG2 hepatic cancer cells.
基金Supported by A grant from Foundation of High Level Talented Specialists of Guizhou Province,China,No. TZJF-200850a grant from Foundation of the Program for Tackling Key Problems of Guizhou Science and Technology Department,China,No. 2010GZ97666
文摘AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2(Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group(A);CCl4-induced hepatic fibrosis group(B);blueberry prevention group(C);Dan-shao-hua-xian capsule(DSHX) prevention group(D);and blueberry + DSHX prevention group(E).Liver fibrosis was induced in rats by subcutaneous injection of CCl4 and a high-lipid/low-protein diet for 8 wk(except the control group).The level of hyaluronic acid(HA) and alanine aminotransferase(ALT) in serum was examined.The activity of superoxide dismutase(SOD),glutathione-S-transferase(GST) and malondialdehyde(MDA) in liver homogenates was determined.The degree of hepatic fibrosis was evaluated by hematoxylin and eosin and Masson staining.Expression of Nrf2 and NADPH quinone oxidoreductase 1(Nqo1) was detected by real-time reversed transcribed-polymerase chain reaction,immunohistochemical techniques,and western blotting.RESULTS:Compared with group B,liver indices,levels of serum HA and ALT of groups C,D and E were reduced(liver indices:0.038 ± 0.008,0.036 ± 0.007,0.036 ± 0.005 vs 0.054 ± 0.009,P<0.05;HA:502.33 ± 110.57 ng/mL,524.25 ± 255.42 ng/mL,499.25 ± 198.10 ng/mL vs 828.50 ± 237.83 ng/mL,P<0.05;ALT:149.44 ± 16.51 U/L,136.88 ± 10.07 U/L,127.38 ± 11.03 U/L vs 203.25 ± 31.62 U/L,P<0.05),and SOD level was significantly higher,but MDA level was lower,in liver homogenates(SOD:1.36 ± 0.09 U/mg,1.42 ± 0.13 U/mg,1.50 ± 0.15 U/mg vs 1.08 ± 0.19 U/mg,P<0.05;MDA:0.294 ± 0.026 nmol/mg,0.285 ± 0.025 nmol/mg,0.284 ± 0.028 nmol/mg vs 0.335 ± 0.056 nmol/mg,P<0.05).Meanwhile,the stage of hepatic fibrosis was significantly weakened(P<0.05).Compared with group A,the activity of GST liver homogenates and expression levels of Nrf2 and Nqo1 in group B were elevated(P<0.05).The expression level of Nrf2 and Nqo1 in groups C,D,and E were increased as compared with group B,but the difference was not significant.CONCLUSION:Blueberry has preventive and protective effects on CCl4-induced hepatic fibrosis by reducing hepatocyte injury and lipid peroxidation.However,these effects may not be related to the activation of Nrf2 during long-term of CCl4.
文摘No study has been performed on identifying microRNAs(miRNAs) and their targets in cotton although cotton is one of the most important fiber and economic crops around the world.In this study,
基金supported by the earmarked fund for the China Agriculture Research System(beef)(CARS-37)the Special Fund for Innovation Team of Modern Agricultural Industrial Technology System in Shandong Province,China(SDAIT-09-09)the Funds of Shandong“Double Tops”Program,China(SYL2017XTTD12)
文摘In order to improve beef color and color stability, step-chilling （SC） was applied on excised bovine Iongissimus lumborum muscle, with chilling starting at 0-4℃ for 5 h, then holding the temperature at 12-18℃ for 6 h, followed by 0-4℃ again until 24 h post-mortem, pH and temperature were measured during rigor on SC loins as well as those subjected to routine chilling （RC, 0-4℃, till 24 h post-mortem）. Color L＊, a＊, b＊ values, metmyoglobin （MetMb） content, MetMb reducing ability （MRA） and NADH content were determined on samples aged for 1, 7, and 14 d. Sarcoplasmic proteome analysis was only conducted on d 1 samples. The results showed muscles subjected to SC maintained a temperature at around 15℃ for 5 to 10 h post-mortem, and exhibited a slow temperature decline, but rapid pH decline. Beef steaks treated with SC had higher L＊, a＊, b＊ and chroma values than those of RC samples at 1 and 7 d chilled storage （0-4℃）, while showing no significant difference for a＊, b＊ and chroma values at d 14. The SC samples also exhibited a lower relative content of surface MetMb, higher MRA and NADH content, compared with RC beef steaks during storage, indicating the SC-treated beef showed an improved color stability. Eleven differential protein spots/nine proteins were identified by two-dimensional gel electrophoresis and mass spectrometry, and those proteins were mainly involved in redox, chaperone binding, metabolic and peroxidase activity. Oxidoreductases play a role in decreasing the oxidation-induced myoglobin oxidation and benefiting the production of NADH, and finally improving the colour of beef. Of these, pyruvate dehydrogenase E1 component subunit beta showed a positive correlation with color L＊, a＊, b＊ values and accounted for more than 60% of the variation in color values; this protein can be considered as a potential beef color biomarker. The present study provided valuable information for studies on the molecular mechanism of color improvement from step-chilling, as well as for identifying markers associated with beef color.
基金supported by National Natural Science Foundation of China (81272930, 81322038, 31260208, and U1132605)the Science and Technological Key Project of Yunnan Province (2012FB185)West Light Foundation of the Chinese Academy of Sciences (to R.L.)
文摘The WW domain-containing oxidoreductase（WWOX） is a tumor suppressor in a variety of cancers, including breast cancer. Reduced WWOX expression is associated with the basal-like subtype and a relatively poor disease-free survival rate among breast cancer patients. Though several WWOX partners have been identified, the functional mechanisms of WWOX＇s role in cancers have not been fully addressed to date. In the current study, we found WWOX suppresses expression of KLF5—an oncogenic transcription factor—at protein level, and suppresses cancer cell proliferation in both bladder and breast cancer cell lines. Furthermore, we demonstrated that WWOX physically interacts with KLF5 via the former＇s WW domains and the latter＇s PY motifs. Interestingly, we found the expression of WWOX negatively correlates with KLF5 expression in a panel of breast cancer cell lines. Taken together, we conjecture that WWOX may suppress cancer cell proliferation partially by reducing the expression of KLF5.
基金We acknowledge Higher Education Commission (HEC), Islamabad, Pakistan, for providing financial support through International Research Support Initiative Program (IRSIP) to conduct this research at University of Sydney, Australia.
文摘Glyphosate is an important organophosphonate herbicide used to eliminate grasses and herbaceous plants in many vegetation management situations.Its extensive use is causing environmental pollution,and consequently,there is a need to remove it from the environment using an eco-friendly and cost-effective method.As a step to address this problem,a novel bacterial strain Comamonas odontotermitis F2,capable to utilize glyphosate as a carbon(C)and/or phosphorus(P)source,was isolated from a glyphostate-contaminated field soil in Australia and characterized.Response surface methodology(RSM)employing a 2^3 full factorial central composite design was used to optimize glyphosate degradation by C.odontotermitis P2 under various culture conditions.The strain C.odontotermitis P2 was proficient in degrading 1.5 g L^-1 glyphosate completely within 104 h.The optimal conditions for the degradation of glyphosate were found to be pH 7.4,29.9℃,and an inoculum density of 0.54 g L^-1,resulting in a maximum degradation of 90%.Sequencing of glyphosate oxidoreductase(GOX)and C-P lyase(phnJ)genes from C.odontotermitis P2 revealed 999c and 93%identities to already reported bacterial GOX and phnJ genes,respectively.The presence of these two genes in C.odontotermitis indicates its potential to degrade glyphosate through GOX and C-P lyase metabolic pathways.This study demonstrates the potential of C.odontotermitis P2 for efficient degradation of glyphosate,which can be exploited for remediation of glyphosate.
